A. Field of Invention
The invention pertains to a method for the quantitative assay of multiple analytes. In particular, the invention pertains to the quantitative affinity assay of multiple analytes based on the specific binding of analytes to their binding partners.
B. Description of the Related Art
For some time now, affinity assays have been used to determine the quantity of chemical and biochemical compounds (analytes). Such tests may include the use of fluorescence immunoassays as well as fluorescence immunosensors.
Document DE 196 28 002 has established methods for the simultaneous assay of multiple analytes. Analyte specific antibodies are used for a sandwich assay. According to specificity, the antibodies are immobilized separately and consecutively in different regions within a range of exposure.
The sample with the analytes and differently labeled, analyte specific antibodies are placed in a container. The number of the antibodies must equal the number of analytes in the assay. The different analyte specific labeled antibodies bind to different locations within the exposure range, dependent on the localization of the earlier immobilized analyte specific antibodies. The quantity of the analytes is determined by time- and space-resolved fluorimetry.
It follows that in this procedure each analyte requires exactly one analyte specific antibody and one immobilized antibody, which itself also has to be analyte specific. As a rule, the use of different labeled antibodies necessitates different excitation wavelengths or space-resolved measurements. Accordingly, the setup for the measurement has to be complex.
In this context, the different labeled antibodies are either different antibodies labeled with different dyes or, alternatively, different antibodies labeled with identical dyes.